UA68937A - Method for treating cutaneous and muscular wounds - Google Patents

Abstract

The method for treating cutaneous and muscular wounds provides for grafting the suspension of cultured cells of human connective tissue, namely cryopreserved embryonic mesenchymal cells to the wound area.

Description

Description of the invention

The invention belongs to the field of medicine, namely combustiology and plastic surgery and can be used for the treatment of large superficial and deep burns, frostbite, donor and scalped wounds, necrotic vasculitis, trophic ulcers of various genesis, etc.

There is a well-known traditional method of treating skin and muscle wounds by closing them with autologous skin mesh grafts (1).

The main disadvantages of this method are the need for significant resources of autoplastic material (reliable and complete healing occurs only with plasticity ratios of 1:2-1:3) and its traumatic nature. In addition, complications in the form of the formation of keloid scars and contractures are possible.

The method closest to the claimed method is the method of treating burn skin-muscle wounds, according to which isolated adult allofibroblasts, which are connective tissue cells, after cultivation are applied to the wound surface in the form of a cell suspension or a cell monolayer with seeded 72 doses 200-50 Otis. kl./ml II.

The main drawback of this method is the insufficiently fast healing of wounds, which is associated with a relatively low engraftment of fibroblasts of the aloskin of an adult and a limited period of their existence in the wound due to their antigenicity and sensitivity to environmental factors. In addition, when the wound healing process is disturbed, due to the inhibition of the growth of own keratinocytes under the action of allofibroblasts, the development of complications related to the formation of keloid scars is possible.

The invention is based on the task of improving the method of treating skin-muscle wounds by using a transplant material with a wider range of biological properties and, thus, achieving a reduction in the healing time of skin-muscle wounds while simultaneously reducing the likelihood of complications in the form of keloid scars.

This task is solved by the fact that in the method of treating skin and muscle wounds by transplanting into the area of the "defect" a suspension of cultured human connective tissue cells, according to the invention, as connective tissue cells use cryopreserved mesenchymal cells of a human embryo (Meso cells), which are transplanted by applying to the surface wounds or inoculations in the thickness of the granulation tissue. -

Mesocells are obtained from mesenchymal-mesodermal tissues of a human embryo. They are characterized by a high content of stem cells and have significant resistance to environmental factors, plasticity, low antigenicity and the ability to produce a large number of growth factors and cytokines. at

Such biological properties allow these cells to take root and function long-term in the wound. with

Mesocells are capable of actively proliferating and differentiating during transplantation, as well as creating favorable conditions for the proliferation of autokeratinocytes in the wound and specific differentiation of other types of cells (keratinocytes, myocytes, chondrocytes, fibroblasts, adipocytes, etc.).

Thanks to these features, Mesocells during transplantation are able to correct epithelial-mesenchymal relations in the wound, which actively influence the wound healing process, shortening its duration, and provide "healing of the skin-muscular wound without pathological scarring." C 50 The use of cryopreserved Mesocells excludes the possibility of wound infection due to the transplant, because in the process of preserving the Mesocells, they are examined in accordance with the existing legislation.

The method is carried out as follows.

In the operating room, under local or general anesthesia, after toileting and special treatment of the wound surface, pre-thawed and cultured Mesocells, in the form of a cell suspension b (200-500 cells/ml), depending on the area and localization of the wound defect, are applied to wound surface or

Ge | inoculated into the thickness of the granulation tissue, after which the wound is closed with a biological or non-adhesive aseptic coating. ее, Clinical examples со 20 Example 1

Patient G., 19 years old. Diagnosis: Necrotic vasculitis. Necrosis of soft tissues of the upper and lower limbs on t area of 79o of the body surface.

The diagnosis was established 5 years ago. For the purpose of surgical treatment of necrotic defects of the skin and soft tissues, the patient came to the clinic several times. Autodermoplasty was accompanied by complicated 29 engraftment of autografts in the form of lysis of transplanted flaps on 50-6095 square meters of closed wounds, i.e. in required repeated plastics. A shortage of donor resources of intact skin has developed.

At the time of admission to the hospital, the granulating wounds of the upper limbs had an area of 395, and the skin deficiency on the lower limbs amounted to 490 of the body surface. In order to overcome the shortage of plastic material and improve the engraftment of skin autoclaves, it was decided to combine autodermoplasty with the transplantation of 60 cryopreserved Mesocells.

The transplant was prepared from cultured cryopreserved mesocells of the second passage. On the upper limbs, only cryopreserved Mesocells were transplanted, which were inoculated with the help of a syringe into the thickness of the granulations with a frequency of one injection per 2cm 2, the seeding dose was 25 Otis. cl/ml of suspension on the lower - autodermoplasty with a mesh skin flap was combined with the application of a suspension of cryopreserved mesocells on the wound surface in the gaps of the skin flap.

On the 7th day after the transplantation operation, marginal epithelization with a width of about 2 cm was markedly expressed on the wounds of the upper extremities, and multiple areas of islet epithelization of cells appeared from the place of transplant insertion in the granulation layer. On the wounds of the lower extremities, pronounced marginal epithelization was observed in the interstices of mesh autografts.

On the 14th day, the wounds on the lower limbs were practically epithelialized, and on the upper limbs their area was reduced by half. On the 18th day after plastic surgery, the wounds on the upper limbs were completely epithelialized.

Example 2

Patient S. 45 years old. Diagnosis: ShB-1U flame burns of the torso, upper and lower limbs on an area of 2090 7/o of the body surface; damage to the respiratory tract; burn shock Severe course of burn disease.

To free the burn wounds from necrosis on the 4th day, a primary necrotomy was performed on the back and right shoulder on the area of 1295 of the body surface. After the removal of necrosis, the wound surfaces were closed with split autografts with a plasticity ratio of 1:6, which required 2905 autoskin.

On the first dressing after autodermoplasty, a culture of cryopreserved 7/5 Mesocells of the third passage was transplanted at a seed dose of 0.000 mg. kl/"ml by applying to the surface of the wounds in the interstices of the mesh graft, after which the wounds were covered with a substrate film on which the cells were cultured.

On the dressing after 7 days (the 14th day after receiving the injury), almost complete epithelialization of the interstices of the mesh grafts was noted. On the 16th day, the patient was left without bandages, because the surfaces of the wounds were completely epithelialized.

When viewed through BMS. noted restored full-fledged skin without dense scars and contractures.

Example C

Patient T, 36 years old. Diagnosis: Keloid scar of the front chest wall, which appeared 2.5 years ago after a boil.

Previously, the keloid was excised three times with the use of triamcinalone in the interoperative period. The last surgical treatment was cryodestruction of the scar 14 months ago. At the time of admission to the hospital, there was a keloid on the skin of the patient's chest with uneven edges, up to 2 cm high, with a crusty surface, with a maximum diameter of about 5 cm.

In the operating room, under general anesthesia, the keloid was excised within healthy tissues and hemostasis was performed. A transplant of cryopreserved Mesocells of the second passage was applied to the bottom and edges of the wound. M zo From above, the transplant was covered with a split skin flap taken from the anterolateral surface of the left thigh, and the edges of the flap overlapped the healthy skin by 1 cm. The first bandage was made on the 6th day, it was established that the autograft is tightly fixed to the wound, it is warm, pink over the entire area. The postoperative course was smooth. The patient was examined a year later, there was no recurrence of keloid.

Example 4 so

Patient V., 22 years old. Diagnosis: Burns of ShB-1U art. both lower extremities to 3595 surface. bodies Burn shock. Ge

On the 3rd day after elimination of burn shock, the patient underwent a primary necroctomy in the operating room. The right lower extremity was covered with split autografts combined with xenografts.

Transplants of cryopreserved mesocells of the 2nd and 3rd passages were applied to the left lower leg.

Cultivation, short-term cultures of cryopreserved Mesocells were used on the left thigh. with c Transplants are covered with mesh autografts with a plastic ratio of 1:8-1:10. From above, the wounds are covered with xenoskin and a non-stick bandage. ;" On the 7th day, the first bandage was made. On the right leg there were areas of lysis of autografts, accumulation of serous and hemorrhagic fluid under them. On the left leg, the bandages were removed and replaced with xenoskin; which was not touched, the wounds were moderately moist with a white coating, painless. On the 12th day, a spontaneous one occurred

Ge" epithelization of the gaps of autografts, that is, elimination of wounds. On the right limb, a similar picture was achieved on the 19th day, and the loss of autografts as a result of their lysis amounted to 9-1195, which required additional autodermoplasty.

Ge) During the examination after b months, the restored skin on the left lower limb was noted, soft, elastic, without the formation of scars and contractures. 19 patients hospitalized in the Kharkiv Burn Center with "M" for burns, frostbite, trophic and post-traumatic skin defects of various areas, depths and localizations were treated with the claimed method.

The effectiveness of the treatment was evaluated by the nature of the wound process and the time of wound healing. In the dynamics of observation, microbiological and cytological studies were carried out. Not a single case of complications related to the transplantation of cryopreserved Mesocells was recorded.

As early as 3-4 days after transplantation, a suspension of cryopreserved mesocells was observed in patients

P» normalization of the character of cytograms of wound prints according to the regenerative type.

The table shows the results of treatment using cryopreserved Mesocells in comparison with the prototype and the traditional method. 60 5

It can be seen from the table that after treatment with Mesocells, the healing of both superficial and deep injuries of the skin and underlying tissues occurs earlier than after treatment with a traditional method or a prototype method. -

Sources of information: 1. Paramonov B.A., Porembskyi J.O., Yablonskyi V.G. Burns: Guide for doctors. - St. Petersburg: SpetsLit, 2000. - p. 244-247. 2. Fedorov V.D., Sarkysov D.S., Alekseev A.A. The use of fibroblast cultures for treatment / chronic inflammation. -Annal of Chir., 1996. -Mo4. - p. 16-20.

Claims (2)

The formula of the invention 1. The method of treating skin and muscle wounds by transplanting cultured human connective tissue cells into the area of the suspension defect, which differs in that cryopreserved mesenchymal cells of a human embryo are used as connective tissue cells. 2. The method of treating skin and muscle wounds according to claim 1, which is characterized by the fact that the transplantation of cryopreserved mesenchymal cells of a human embryo is carried out by application to the surface of the wound or inoculation in the thickness of the granulation tissue. Official bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated microcircuits", 2004, M 8, 15.08.2004. State Department of Intellectual Property of the Ministry of Education and Science of Ukraine. « cha (ze) (o) (ee) (Se) - with . and? (o) (ee) se) (95) that 60 b5